This study compared the morphological and molecular characteristics of the two Pampus spp. in Korea, Japan and China to clarify the long-confused taxonomic status of the two Korean pomfrets named as "Byeong-eo" and "Deok-dae". Among them "Byeong-eo" was identified as Pampus punctatissimus in having the following morphological traits: patch of wavy ridges extending beyond the anterior tip of the pectoral fin, the groove on the lower ridge of gill cover absent, and the number of vertebrae 34. On the other hand, "Deok-dae" was identified as Pampus argenteus in having the following morphological traits: patch of wavy ridges not reaching to the anterior tip of the pectoral fin, the groove on the lower ridge of gill cover reaching to the lower jaw, and the number of vertebrae 38~42. In addition, "Byeong-eo" almost corresponded to P. punctatissimus from Japan in the mitochondrial DNA cytochrome c oxidase subunit I sequences (genetic distance=0.1%), while "Deokdae" showed genetic distance of 0.3% when compared to P. argenteus from China. Our comprehensive results suggest the validify of P. punctatissimus for "Byeong-eo", and P. argenteus for "Deok-dae".
Purpose: Due to the many problems with commercially available vaccines, the production of effective vaccines against brucellosis is a necessity. The aim of this study was to evaluate the immune responses caused by the chimeric protein consisting of trigger factor, Bp26, and Omp31 (TBO) along with aluminum hydroxide (AH/TBO) and selenium (Se/TBO) nanoparticles (NPs) as adjuvants in mouse model. Materials and Methods: Recombinant antigen expression was induced in Escherichia coli BL21 (DE3) bacteria using IPTG (isopropyl-d-1-thiogalactopyranoside). Purification and characterization of recombinant protein was conducted through NiFe3O4 NPs, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Western blot. NP characteristics, including morphology and particle size, were measured in vitro. The recombinant TBO was loaded on to AH and Se NPs and were administered subcutaneously. After mice immunization, measurement of antibody titter and protection assay was performed. Results: The average sizes of AH and Se NPs were about 60 nm and 150 nm, respectively. The enzyme-linked immunosorbent assay results showed that the serum of mice immunized by subcutaneous injection with both nanovaccines produced significant immunoglobulin G (IgG) responses against the chimeric antigen. The results of TBO-specific IgG isotype (IgG2a/IgG1) analysis showed that both AH and Se NPs induced a type to T-helper immune response. In addition, the results of the challenge with the pathogenic strain of Brucella melitensis 16M showed that vaccinated mice with AH/TBO NPs indicated a higher reduction of bacterial culture than immunized mice with Se/TBO NPs and TBO alone. Conclusion: The results showed that AH NPs carrying chimeric antigen can be a promising vaccine candidate against brucellosis by producing protective immunity.
Hala Khyami-Horani;Amal Al-Aboudi;Musa Abu Zarga;Monther Sadder;Halima Othman
Microbiology and Biotechnology Letters
/
v.51
no.4
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pp.474-483
/
2023
Members of the genus Streptomyces produce more than 70% of antibiotics. The rise in antibiotic resistance globally enhanced the search for novel species with the ability to produce new bioactive compounds. This study was initiated to investigate different regions in Jordan for previously uncultured and rare Streptomyces species capable of producing novel antimicrobial compounds especially active against bacteria resistant to antibiotics. A total of 191 Streptomyces strains were isolated from 26 soil samples collected from different geographic regions in Jordan. Isolates were characterized based on colony and cellular morphology as well as using 16S rRNA gene sequencing. These isolates were screened for their ability to produce antibiotics by the perpendicular-cross streak method, and then tested by well diffusion method against tested pathogens. Fifty-four isolates showed potential to produce antimicrobial products especially active against resistant bacteria, 20.1% of the isolates showed inhibitory effect against Staphylococcus aureus, 16.9% against clinical MSSA strains, and 18.0% against MRSA: whereas only 4.2% against Esherichia coli, 3.2% against Klebsiella pneumonia, 2.7% against Pseudomonas aeruginosa, and 10.0% against clinical Candida albicans. Three isolates were selected for further identification due to their antibacterial activity against S. aureus, MRSA, and MSSA. These isolates were identified as follows; Streptomyces aburaviensis DSa3, Streptomyces alboniger SAb7 and Streptomyces misionensis ZAb2, based on cultural, biochemical characteristics and molecular analysis of the 16S rRNA.
Jegadeesh Raman;Young-Joon Ko;Jeong-Seon Kim;Da-Hye Kim;Soo-Jin Kim
Journal of Microbiology and Biotechnology
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v.34
no.3
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pp.710-724
/
2024
Flavobacterium can synthesize xanthophyll, particularly the pigment zeaxanthin, which has significant economic value in nutrition and pharmaceuticals. Recently, the use of carotenoid biosynthesis by bacteria and yeast fermentation technology has shown to be very efficient and offers significant advantages in large-scale production, cost-effectiveness, and safety. In the present study, JSWR-1 strain capable of producing xanthophyll pigment was isolated from a freshwater reservoir in Wanju-gun, Republic of Korea. Based on the morphological, physiological, and molecular characteristics, JSWR-1 classified as belonging to the Flavobacterium species. The bacterium is strictly aerobic, Gram-negative, rod-shaped, and psychrophilic. The completed genome sequence of the strain Flavobacterium sp. JSWR-1 is predicted to be a single circular 3,425,829-bp chromosome with a G+C content of 35.2% and 2,941 protein-coding genes. The optimization of carotenoid production was achieved by small-scale cultivation, resulting in zeaxanthin being identified as the predominant carotenoid pigment. The enhancement of zeaxanthin biosynthesis by applying different light-irradiation, variations in pH and temperature, and adding carbon and nitrogen supplies to the growth medium. A significant increase in intracellular zeaxanthin concentrations was also recorded during fed-batch fermentation achieving a maximum of 16.69 ± 0.71 mg/l, corresponding to a product yield of 4.05 ± 0.15 mg zeaxanthin per gram cell dry weight. Batch and fed-batch culture extracts exhibit significant antioxidant activity. The results demonstrated that the JSWR-1 strain can potentially serve as a source for zeaxanthin biosynthesis.
Background: Post-ovulatory aging (POA) of oocytes is related to a decrease in the quality and quantity of oocytes caused by aging. Previous studies on the characteristics of POA have investigated injury to early embryonic developmental ability, but no information is available on its effects on mitochondrial fission and mitophagy-related responses. In this study, we aimed to elucidate the molecular mechanisms underlying mitochondrial fission and mitophagy in in vitro maturation (IVM) oocytes and a POA model based on RNA sequencing analysis. Methods: The POA model was obtained through an additional 24 h culture following the IVM of matured oocytes. NMN treatment was administered at a concentration of 25 μM during the oocyte culture process. We conducted MitoTracker staining and Western blot experiments to confirm changes in mitochondrial function between the IVM and POA groups. Additionally, comparative transcriptome analysis was performed to identify differentially expressed genes and associated changes in mitochondrial dynamics between porcine IVM and POA model oocytes. Results: In total, 32 common genes of apoptosis and 42 mitochondrial fission and function uniquely expressed genes were detected (≥ 1.5-fold change) in POA and porcine metaphase II oocytes, respectively. Functional analyses of mitochondrial fission, oxidative stress, mitophagy, autophagy, and cellular apoptosis were observed as the major changes in regulated biological processes for oocyte quality and maturation ability compared with the POA model. Additionally, we revealed that the activation of NAD+ by nicotinamide mononucleotide not only partly improved oocyte quality but also mitochondrial fission and mitophagy activation in the POA porcine model. Conclusions: In summary, our data indicate that mitochondrial fission and function play roles in controlling oxidative stress, mitophagy, and apoptosis during maturation in POA porcine oocytes. Additionally, we found that NAD+ biosynthesis is an important pathway that mediates the effects of DRP1-derived mitochondrial morphology, dynamic balance, and mitophagy in the POA model.
Purpose: Trop family proteins, including epithelial cell adhesion molecule (EpCAM) and Trop-2, have garnered attention as potential therapeutic and diagnostic targets for various malignancies. This study aimed to elucidate the clinicopathological significance of these proteins in gastric carcinoma (GC) and to reinforce their potential as biomarkers for patient stratification in targeted therapies. Materials and Methods: Immunohistochemical (IHC) analyses of EpCAM and Trop-2 were performed on GC and precancerous lesions, following rigorous orthogonal validation of the antibodies to ensure specificity and sensitivity. Results: Strong membranous staining (3+) for Trop-2 was observed in 49.3% of the GC cases, whereas EpCAM was strongly expressed in almost all cases (93.2%), indicating its widespread expression in GC. A high Trop-2 expression level, characterized by an elevated H-score, was significantly associated with intestinal type by Lauren classification, gastric mucin type, presence of lymph node metastasis, human epidermal growth factor receptor 2-positivity, and Epstein-Barr virus (EBV)-positivity. Patients with a high Trop-2 expression level exhibited poorer survival outcomes on univariate and multivariate analyses. High EpCAM expression levels were prevalent in differentiated histologic type, microsatellite instability-high, and EBV-negative cancer, and were correlated with high densities of CD3 and CD8 T cells and elevated combined positive score for programmed death-ligand 1. Conclusions: These results highlight the differential expression of Trop-2 and EpCAM and their prognostic implications in GC. The use of meticulously validated antibodies ensured the reliability of our IHC data, thereby offering a robust foundation for future therapeutic strategies targeting Trop family members in GC.
Jeongwoo Shin;Sangki Choi;Heejong Son;Byungryul An
Journal of Korean Society of Water and Wastewater
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v.38
no.4
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pp.199-208
/
2024
In this paper, the adsorption removal characteristic for 10 species of perfluoroalkyl and polyfluoroalkyl substances (PFAS) was investigated using GAC and modified GAC (GAC-Cu). After modification with Cu(II), the amount of copper was to 1.93 and 4.73 mg/g for GAC and GAC-Cu, respectively. The total amount of 10 species of PFAS per specific area was obtained to 0.548 and 0.612 ng/m2 for GAC and GAC-Cu, respectively. A series of batch test confirmed lower efficiency was observed with a smaller number of carbon chain length and the removal efficiency of PFCA (perfluoroalkyl carboxylic acids) was lower than that of PFSA (perfluoroalkyl sulfonic acids) with the same carbon chain length. Regarding the pH effect, the adsorption capacity was decreased with increase of pH due to the increase of electrostatic repulsion. According to pseudo first and second order (PFO and PSO) kinetic models, while the values of equilibrium uptake and time did not show significant difference, a difference in uptake was observed between 24-48h. Furthermore, based on correlation analysis, Log Kow and uptake have a high correlation with molecular weight (M.W.) and initial concentration, respectively. These results show that long-chain PFAS have higher removal efficiency due to their increased hydrophobicity.
Jeong-Byoung Chae;Seung-Uk Shin;Serim Kim;Hansong Chae;Won Gyeong Kim;Joon-Seok Chae;Hyuk Song;Jung-Won Kang
Journal of Veterinary Science
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v.25
no.5
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pp.59.1-59.10
/
2024
Importance: Despite advancements in herd management, feeding, and pharmaceutical interventions, neonatal calf diarrhea (NCD) remains a major global health concern. Bacteria, viruses, and parasites are the major contributors to NCD. Although several pathogens have been identified in the Republic of Korea (ROK), the etiological agents of numerous NCD cases have not been identified. Objective: To identify, for the first time, the prevalence and impact of Boosepivirus (BooV) on calf diarrhea in the ROK. Methods: Here, the unknown cause of calf diarrhea was determined using metagenomics We then explored the prevalence of certain pathogens, including BooV, that cause NCD. Seventy diarrheal fecal samples from Hanwoo (Bos taurus coreanae) calves were analyzed using reverse transcriptase and quantitative real-time polymerase chain reaction for pathogen detection and BooV isolate sequencing. Results: The complete genome of BooV was detected from unknown causes of calf diarrhea. And also, BooV was the most frequently detected pathogen (35.7%) among 8 pathogens in 70 diarrheic feces from Hanwoo calves. Co-infection analyses indicated that most BooV-positive samples were solely infected with BooV, indicating its significance in NCD in the ROK. All isolates were classified as BooV B in phylogenetic analysis. Conclusions and Relevance: This is the first study to determine the prevalence and molecular characteristics of BooV in calf diarrhea in the ROK, highlighting the potential importance of BooV as a causative agent of calf diarrhea and highlighting the need for further research on its epidemiology and pathogenicity.
Although Sumunmulbangdui wetland at the Halla Mountain in Jeju Island, a kind of montane wetlands, has been considered to bear high biodiversity, no study has been reported on the bacterial diversity. In this study, soil and water samples were collected from the wetland in order to isolate novel bacterial species. Bacterial strains belonging to the phyla Bacteroidetes, Firmicutes, and Actinobacteria were isolated after spreading soil and water samples onto solid agar media. The 16S rRNA gene sequences of the strains assigned to the three phyla were compared to those of type strains of the species in the phyla. The strains that showed less than 98.7% 16S rRNA gene sequence similarity to the validly published species were considered to be novel species candidates. A total of 32 strains were regarded as novel species candidates in the phyla Bacteroidetes, Firmicutes, and Actinobacteria. Diversity of novel species candidates was very low; the candidates were confined to only few genera. In the Bacteroidetes, 13 novel candidate species were affiliated with the genera Mucilaginibacter, Sphingobacterium, Pedobacter, Flavobacterium, and Chryseobacterium. A total of 13 novel candidate species that assigned to the genera Paenibacillus Lysinibacillus, and Bacillus were identified in the phylum Firmicutes. Only two candidate species that belonged to the genera Mycobacterium and Nocardia were excavated in the Actinobacteria. Cultural, physiological, and chemotaxonomic characteristics have been determined for the novel species candidates, and the characteristics are described in this study.
Chinese cabbage ('Baechu') Kimchi was fermented at the three different temperatures right after it was prepared. Samples were taken everyday for measuring bacterial populations, pH, and titratable acidity through the whole periods of fermentation up to 50 days. pH values and developed acidity were significantly affected by the fermenting temperatures of 4, 10, and $20^{\circ}C$, suggesting that different bacterial flora has been established by the temperatures exposed. The modified MRS agar containing vancomycin (300 $\mu$g/mL) was used for isolating the vancomycin-resistant LAB strains and 127 isolates were finally obtained. Of the LAB isolates, 13 isolates were subjected to the identification experiments based on the biochemical characteristics and the molecular-typing approach, an ITS-PCR, whether they belong to the genus Leuconostoc or not. The data obtained from API 50 CHL kit resulted that six isolates were identified as the members of Leuconostoc and six as Lactobacillus brevis strains except for a single isolate YKI 30-0401, which was not able to be identified because its biochemical traits were not matched to the database of API 50 CHL kit. It was noted that some isolates were distinct in a couple of some biochemical characteristics compared with those of the reference Leuconostoc species. To overcome the limitations experienced in the commercial identification products above, an ITS-PCR experiment was also conducted for the isolates, resulting that eight isolates belong to Leu. mesenteroides ssp. mesenteroides or dextranicum with a single band of 564 bp, and four to L. brevis strains. The ITS-PCR profiles clearly differentiated the closely-related LAB isolates for which same results were obtained by the biochemical method. This molecular approach, however, failed to produce the amplicons for the YKI 20-1003, leaving the strain unidentified. Judging from the identification data obtained in the Kimchi fermented at $4^{\circ}C$ or $10^{\circ}C$, Leuconostoc spp. including Leu. mesenteroides/dextranicum were likely predominant species in the earlier stage and L. brevis occurred at the high level through the whole period. By contrast, L. brevis, as one of the major flora, possibly lead the fermentation from the beginning in the Kimchi fermented at $20^{\circ}C$.}C$.
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